光干涉生物亲和性检测系统设计
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摘要
分子互作检测是目前食品安全检测、临床癌症病理筛查等检测领域的研究热点。为了对生物分子间相互作用进行实时、快速检测,本文以光干涉技术为理论基础,设计了基于光纤生物传感器的分子互作检测系统。采用了石英光纤与自聚焦透镜耦合结构,提高了光源与干涉光的传输光路耦合效率。采用STM32微处理器作为主控制器,实现了数据采集、三维传动机构控制、恒温震荡控制、通讯等功能。最后,采用所设计的分子互作检测系统及美国Fortebio公司的BLItz分子互作检测系统,分别进行不同浓度的定量检测实验,实时检测IgG与抗原分子蛋白A的结合与解离反应。实验结果表明:本文设计的系统检测极限达到10μg/mL,重复性CV值小于6%,与BLItz接近。所设计系统检测的自动化程度高、检测过程中无需清洗、不产生交叉污染、成本低,能够满足药代动力学研究需要。
Molecular interaction detection is a hot topic in the fields of food safety detection,clinical cancer pathological screening,and other areas of research.To realize real-time and rapid detection of interactions between biomolecules,a molecular interaction detection system based on a fiber biosensor was designed in this study.The coupling structure of a self-focusing lens and quartz fiber was adopted to improve the coupling efficiency of optical transmission between the light source and interference light.A STM32 microprocessor was used as the main controller to realize data acquisition,3 D transmission mechanism control,constant temperature oscillation control,and other functions.Finally,the designed and BLItz molecular interaction detection systems were used to detect the different concentrations of immunoglobulin IgG and protein A,respectively.The binding and dissociation reactions of the IgG molecule and antigen molecule protein A were detected in real time.The experimental results show that the detection limit of the system could reach 10μg/mL,and the repeatability CV value was less than 6%,which was close to that of the BLItz.The designed system has the advantages of high automation,no need for cleaning,no cross contamination in the detection process,and low cost.Therefore,it can meet the needs of pharmacokinetics research.
Molecular interaction detection is a hot topic in the fields of food safety detection,clinical cancer pathological screening,and other areas of research.To realize real-time and rapid detection of interactions between biomolecules,a molecular interaction detection system based on a fiber biosensor was designed in this study.The coupling structure of a self-focusing lens and quartz fiber was adopted to improve the coupling efficiency of optical transmission between the light source and interference light.A STM32 microprocessor was used as the main controller to realize data acquisition,3 D transmission mechanism control,constant temperature oscillation control,and other functions.Finally,the designed and BLItz molecular interaction detection systems were used to detect the different concentrations of immunoglobulin IgG and protein A,respectively.The binding and dissociation reactions of the IgG molecule and antigen molecule protein A were detected in real time.The experimental results show that the detection limit of the system could reach 10μg/mL,and the repeatability CV value was less than 6%,which was close to that of the BLItz.The designed system has the advantages of high automation,no need for cleaning,no cross contamination in the detection process,and low cost.Therefore,it can meet the needs of pharmacokinetics research.
引文
[1]POLLET J,DELPORT F,JANSSEN P F,et al..Fiber optic SPR biosensing of DNA hybridization and DNA-protein interactions[J].Biosensors &Bioelectronics,2009,25(4):864-869.
[2]郭团.等离子体共振光纤光栅生物传感器综述[J].光学学报,2018,38(3):75-95.GUO T.Review on plasmonic optical fiber grationg biosensors[J].Acta Optica Sinica,2018,38(3):75-95.(in Chinese)
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[4]JOY C,KRISTA W,CHARLES W,et al..Labelfree detection of biomolecular interactions using Biolayer interferometry for kinetic characterization[J].Silico Biology,2009,5(4):407-413.
[5]LI ZH Y,LIAO CH R,CHEN D N,et al..Labelfree detection of bovine serum albumin based on an in-fiber Mach-Zehnder interferometric biosensor[J].Optics Express,2017,25(15):17105-17113.
[6]薛孟伟.反射式光纤SPR生物传感器的硏究及测试实验[D].济南:山东大学,2017.XUE M W.Research and Experiment of Reflective Fiber SPR Biosensor[D].Jinan:Shandong university,2017.(in Chinese)
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[8]王弋嘉,张崇磊,王蓉,等.差分干涉表面等离子体共振传感器的优化与验证[J].光学精密工程,2013,21(3):672-679.WANG G J,ZHANG CH L,WANG R,et al..Optimization and validation of differential interferometric surface plasmon resonance sensor[J].Opt.Precision Eng.,2013,21(3):672-679.(in Chinese)
[9]衡磊,王冲,钱卫珠,等.生物薄膜干涉技术和ELISA法检测抗体药物免疫原性的方法学比较[J].现代免疫学,2012,32(4):282-286.HENG L,WANG CH,QIAN W ZH,et al..Methodological comparison of ELISA and BLI for the quantitation of anti-drug antibodies in human serum[J].Current Immunology,2012,32(4):282-286.(in Chinese)
[10]童凯,党鹏,汪梅婷,等.采用TiO2薄膜增强光子晶体光纤表面等离子体共振生物传感器灵敏度的建模分析[J].中国激光,2018,45(6):0610002.TONG K,DANG P,WANG M T,et al..Enhancement of sensitivity of photonic crystal fiber surface plasmon resonance biosensor using TiO2film[J].Chinese Journal of Lasers,2018,45(6):0610002.(in Chinese)
[11]MARAGOS C M.Signal amplification using colloidal gold in a biolayer interferometry-based immunosensor for the mycotoxin deoxynivalenol[J].Food Additives&Contaminants:Part A Chemistry,Analysis,Control,Exposure&Risk Assessment,2012,29(7):1108-1117.
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[13]DYSINGER M,KING L E.Practical quantitative and kinetic applications of bio-layer interferometry for toxicokinetic analysis of a monoclonal antibody therapeutic[J].Journal of Immunological Methods,2012,379(1-2):30-41.
[14]史健松,于源华,王美娇,等.光纤生物传感器在HER3抗体药物定量检测中的应用[J].中国光学,2018,11(3):503-512.SHI J S,YU Y H,WANG M J,et al..Application of optical fiber biosensor in quantitative detection of HER3antibody[J].Chinese Optics,2018,11(3):503-512.(in Chinese)
[15]SA J I,CHEN Y P,ZHANG G ZH.Gas cell based on cascaded GRIN lens for optical fiber gas sensor-art[J].Proceedings of SPIE-The International Society for Optical Engineering,2007,12(3):231-237.
[16]王驰,许婷婷,毕书博,等.测量自聚焦光纤透镜聚焦常数的曲线拟合算法[J].光学精密工程,2015,23(12):3309-3315.WANG CH,XU T T,BI SH B,et al..curve-fitting algorithm measuring focusing constant of gradient-index fiber lens[J].Opt.Precision Eng.,2015,23(12):3309-3315.(in Chinese)
[17]卞海溢,高万荣,廖九零.内窥式扫频光源光学相干层析系统的探头设计[J].光子学报,2016,45(9):0911001.BIAN H Y,GAO W R,LIAO J L.Design of the probe of swept source optical coherence tomography for endoscope imaging[J].Acta Photonica Sinica,2016,45(9):0911001.(in Chinese)
[18]丁力,宋志平,徐萌萌,等.基于STM32的嵌入式测控系统设计[J].中南大学学报(自然科学版),2013,44(1):260-265.DING L,SONG ZH P,XU M M,et al..Design of embedded measurement and control system based on STM32[J].Journal of Central South University(Science and Technology),2013,44(1):260-265.(in Chinese)
[19]翟少雄,王长胜,谭立,等.空间高精度扫描伺服系统的驱动控制[J].光学精密工程,2016,24(5):1105-1111.ZHAI SH X,WANG CH SH,TAN L,et al..Driving and controlling of high accuracy scanning servo-system in space[J].Opt.Precision Eng.,2016,24(5):1105-1111.(in Chinese)
[2]郭团.等离子体共振光纤光栅生物传感器综述[J].光学学报,2018,38(3):75-95.GUO T.Review on plasmonic optical fiber grationg biosensors[J].Acta Optica Sinica,2018,38(3):75-95.(in Chinese)
[3]POLLET J,DELPORT F,JANSSEN K P F,et al..Fiber optic SPR biosensing of DNA hybridization and DNA-protein interactions[J].Biosensors and Bioelectronics,2009,25(4):864-869.
[4]JOY C,KRISTA W,CHARLES W,et al..Labelfree detection of biomolecular interactions using Biolayer interferometry for kinetic characterization[J].Silico Biology,2009,5(4):407-413.
[5]LI ZH Y,LIAO CH R,CHEN D N,et al..Labelfree detection of bovine serum albumin based on an in-fiber Mach-Zehnder interferometric biosensor[J].Optics Express,2017,25(15):17105-17113.
[6]薛孟伟.反射式光纤SPR生物传感器的硏究及测试实验[D].济南:山东大学,2017.XUE M W.Research and Experiment of Reflective Fiber SPR Biosensor[D].Jinan:Shandong university,2017.(in Chinese)
[7]唐春晓,李恩邦,王长乐,等.基于多模干涉的反射式光纤生物传感器的研究[J].光电子激光,2011,22(8):1138-1142.TANG C X,LI E B,WANG CH L,et al..Reflection-type fiber-optic biosensor based on multimode interference[J].Journal of Optoelectronics.Laser,2011,22(8):1138-1142.(in Chinese)
[8]王弋嘉,张崇磊,王蓉,等.差分干涉表面等离子体共振传感器的优化与验证[J].光学精密工程,2013,21(3):672-679.WANG G J,ZHANG CH L,WANG R,et al..Optimization and validation of differential interferometric surface plasmon resonance sensor[J].Opt.Precision Eng.,2013,21(3):672-679.(in Chinese)
[9]衡磊,王冲,钱卫珠,等.生物薄膜干涉技术和ELISA法检测抗体药物免疫原性的方法学比较[J].现代免疫学,2012,32(4):282-286.HENG L,WANG CH,QIAN W ZH,et al..Methodological comparison of ELISA and BLI for the quantitation of anti-drug antibodies in human serum[J].Current Immunology,2012,32(4):282-286.(in Chinese)
[10]童凯,党鹏,汪梅婷,等.采用TiO2薄膜增强光子晶体光纤表面等离子体共振生物传感器灵敏度的建模分析[J].中国激光,2018,45(6):0610002.TONG K,DANG P,WANG M T,et al..Enhancement of sensitivity of photonic crystal fiber surface plasmon resonance biosensor using TiO2film[J].Chinese Journal of Lasers,2018,45(6):0610002.(in Chinese)
[11]MARAGOS C M.Signal amplification using colloidal gold in a biolayer interferometry-based immunosensor for the mycotoxin deoxynivalenol[J].Food Additives&Contaminants:Part A Chemistry,Analysis,Control,Exposure&Risk Assessment,2012,29(7):1108-1117.
[12]MCGRATHT F,CAMPBELL K,FODEY T L,et al..An evaluation of the capability of a biolayer interferometry biosensor to detect low-molecularweight food contaminants[J].Analytical and Bioanalytical Chemistry,2013,405(8):2535-2544.
[13]DYSINGER M,KING L E.Practical quantitative and kinetic applications of bio-layer interferometry for toxicokinetic analysis of a monoclonal antibody therapeutic[J].Journal of Immunological Methods,2012,379(1-2):30-41.
[14]史健松,于源华,王美娇,等.光纤生物传感器在HER3抗体药物定量检测中的应用[J].中国光学,2018,11(3):503-512.SHI J S,YU Y H,WANG M J,et al..Application of optical fiber biosensor in quantitative detection of HER3antibody[J].Chinese Optics,2018,11(3):503-512.(in Chinese)
[15]SA J I,CHEN Y P,ZHANG G ZH.Gas cell based on cascaded GRIN lens for optical fiber gas sensor-art[J].Proceedings of SPIE-The International Society for Optical Engineering,2007,12(3):231-237.
[16]王驰,许婷婷,毕书博,等.测量自聚焦光纤透镜聚焦常数的曲线拟合算法[J].光学精密工程,2015,23(12):3309-3315.WANG CH,XU T T,BI SH B,et al..curve-fitting algorithm measuring focusing constant of gradient-index fiber lens[J].Opt.Precision Eng.,2015,23(12):3309-3315.(in Chinese)
[17]卞海溢,高万荣,廖九零.内窥式扫频光源光学相干层析系统的探头设计[J].光子学报,2016,45(9):0911001.BIAN H Y,GAO W R,LIAO J L.Design of the probe of swept source optical coherence tomography for endoscope imaging[J].Acta Photonica Sinica,2016,45(9):0911001.(in Chinese)
[18]丁力,宋志平,徐萌萌,等.基于STM32的嵌入式测控系统设计[J].中南大学学报(自然科学版),2013,44(1):260-265.DING L,SONG ZH P,XU M M,et al..Design of embedded measurement and control system based on STM32[J].Journal of Central South University(Science and Technology),2013,44(1):260-265.(in Chinese)
[19]翟少雄,王长胜,谭立,等.空间高精度扫描伺服系统的驱动控制[J].光学精密工程,2016,24(5):1105-1111.ZHAI SH X,WANG CH SH,TAN L,et al..Driving and controlling of high accuracy scanning servo-system in space[J].Opt.Precision Eng.,2016,24(5):1105-1111.(in Chinese)